Despite decades of research focused mainly on gene transfection, effective intracellular delivery of biologically relevant material remains a difficult task, inhibiting research in a variety of biological and biomedical fields. This proposed work describes a commercially viable intracellular nanomaterial delivery device, the Electrosonic Ejector Microarray (EEM), that is optimized for drug and / or gene / nucleic acid and / or imaging agent delivery into cells via precise control of biophysical action (i.e., concurrent application of sono / mechanoporation, electroporation and thermoporation). The EEM is a novel microelectromechanical systems (MEMS)-enabled device that ejects a sample containing biological cells through microscopic nozzles with incorporated electroporation electrodes, thereby opening pores in the cell membrane via combined mechano / electroporation for uptake of nanomaterials.1 The high ultrasonic frequency of operation and a parallel (array) format enable fast processing of large cell populations at rates between 1 and 100 million cells per second;however, the device can potentially accommodate a wide range of sample sizes, from ~100 nL to arbitrarily large volumes when operated in continuous-flow mode. In addition, the device can be made disposable to eliminate cross-contamination and provides uniform (i.e., identical across an entire cell population) treatment on a single-cell level, which are critical capabilities for sample preparation in clinical applications of cell biology and gene therapy. The EEM is well-suited to basic / applied research, as well as diagnostic and therapeutic uses. Initial investigations will focus on cancer therapies combining mature recombinant protein therapies with emerging RNA interference (RNAi) technologies. The improved understanding of the relationship between the EEM operating parameters and realized bioeffects that will be gained through this proposed work is directly transferable to other application spaces. The primary objective of the proposed work is to develop a commercially viable EEM that demonstrates quantitative performance improvement over currently available nanomaterial delivery technologies. To achieve this objective, (1) an EEM platform that is optimized for cell treatment will be developed, (2) safe treatment of a variety of cell types and characterization of nanomaterial localization will be demonstrated, and (3) the transfection capabilities of the EEM will be optimized and its advantages over traditional transfection technologies quantified. PUBLIC HEALTH RELEVANCE: Development of the Electrosonic Ejector Microarray (EEM) will address the deficiencies of current intracellular drug / gene delivery techniques, which are inhibiting research in a variety of biological and biomedical fields. In particular, successful use of the EEM in development of therapies to treat glioblastomas, which are the most common and lethal brain tumors, will prove its feasibility in a wide range of drug discovery and gene therapy applications.